目的 探索保加利亚乳杆菌GIM1.155对多重耐药质粒（RP4）接合转移的影响。方法 供体菌为含有多重耐药质粒RP4的大肠杆菌 MG1655 K12,具有四环素、卡那霉素、氨苄青霉素三种抗性;受体菌为具有美罗培南抗性的肺炎克雷伯杆菌。供、受体菌菌量及体积比例为 1：1,接合体系为10 ml,最后使用含不同抗生素的抗性平板将供受体菌及接合子筛选出,并计算接合转移频率。探究供、受体菌量梯度(106、107、108、109CFU/ml)以及GIM1.155菌量梯度（107、108、109、1010 CFU/ml）、温度（4、25、37℃）、时间（1、2、4、8、12、24、36、48 h）以及pH值（2、4、6、7、8、9）对RP4质粒接合转移的影响。结果 保加利亚乳杆菌GIM1.155可以促进RP4质粒从大肠杆菌 MG1655 K12到肺炎克雷伯杆菌的接合转移,且促进作用与供受体菌量、GIM1.155菌量以及接合转移的温度、时间、pH有关。结论 一定条件下,保加利亚乳杆菌GIM1.155可促进多重耐药质粒RP4的接合转移。
Abstract
Objective To explore the effect of Lactobacillus bulgaricus GIM1.155 on the conjugative transfer of multi-drug resistant plasmid (RP4). Methods The donor bacterium was Escherichia coli MG1655 K12 containing multi-drug resistant plasmid RP4 with resistance against tetracycline, kanamycin and ampicillin, and the recipient bacterium was Klebsiella pneumoniae with meropenem resistance. The ratio of donor and recipient bacteria was kept at 1:1, and the splicing system was 10mL. Finally, the donor and recipient bacteria and splices were screened out using resistant plates containing different antibiotics, and the frequency of conjugative transfer was calculated. The effects of donor and recipient volume gradients (106, 107, 108, 109 CFUml), as well as GIM1.155 volume gradients (107, 108, 109, 1010 CFU/ml), temperature (4, 25, 37℃), time (1, 2, 4, 8, 12, 24, 36, 48h) and pH (2, 4, 6, 7, 8, 9) on the conjugative transfer of RP4 plasmid were investigated. Results Lactobacillus bulgaricus GIM1.155 could promote the conjugative transfer of RP4 plasmid from E. coli MG1655 K12 to K. pneumoniae, and the promotion effect was dependent on the amount of the donor-recipient bacterium, the amount of the bacterium of GIM1.155, as well as the temperature, time and pH of conjugative transfer. Conclusion Lactobacillus bulgaricus GIM1.155 can promote the conjugative transfer of multi-drug resistant plasmid RP4 under certain conditions.
关键词
保加利亚乳杆菌 /
接合转移 /
多重耐药基因 /
大肠杆菌 /
肺炎克雷伯杆菌
Key words
Lactobacillus bulgaricus /
conjugative transfer /
multidrug resistance gene /
Escherichia coli /
Klebsiella pneumoniae
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